A fluorogenic assay for transketolase from Saccharomyces cerevisiae

In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We exami...

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Bibliographic Details
Published inTetrahedron letters Vol. 44; no. 4; pp. 827 - 830
Main Authors Sevestre, Aurélie, Hélaine, Virgil, Guyot, Ghislain, Martin, Christine, Hecquet, Laurence
Format Journal Article
LanguageEnglish
Published OXFORD Elsevier Ltd 2003
Elsevier
Subjects
Chemistry
Chemistry, Organic
Life Sciences
Physical Sciences
Science & Technology
ENANTIOSELECTIVITY
ALDOLASE
DIRECTED EVOLUTION
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Summary:In order to generate transketolase (TK) with new or improved properties by in vitro evolution, an efficient screening system is an absolute prerequisite for identifying the evolved enzyme variants. We report here an assay allowing us to detect wild type TK activity in vitro by fluorescence. We examined the use of the fluorogenic compound 1 as donor substrate of TK, which is itself non fluorescent but releases a fluorescent product: umbelliferone. Graphic
ISSN:0040-4039
1873-3581
DOI:10.1016/S0040-4039(02)02634-5